The over-arching goal of this Phase I SBIR proposal is to begin to optimize and implement standard reference diets for the Zebrafish (Danio rerio), an animal model of critical importance to the understanding of human health and development of vertebrate organisms. A key problem in the industry is that standardized diets or feed management strategies have not been developed for Danio rerio. These diets are essential to develop rigor and reproducibility in research where zebrafish is the appropriate model. Our solution for the industry, driven by a synergistic collaboration between Meridian Biotech and the laboratory of Dr. Stephen Watts at the University of Alabama at Birmingham, is to develop well-defined open formulation diets for juvenile and adult Zebrafish, that will achieve a desired level of control and whereby use of this modern and effective biomedical animal model is substantially improved. This commercial-academic collaboration is a significant value add to this applied science program. Key ingredients in this formulated diet will include protein packages that contain defined and effective sources, including Single Cell Protein (SCP), and enriched protein source obtained from bacteria or yeast. Diets with these protein sources have been developed in collaboration with Meridian Biotech which have improved shrimp farming feed technologies markedly; thus, we wish to apply this technology to Danio rerio and other relevant models. The over-arching hypothesis of this SBIR proposal is that specific nutrients, provided through selected ingredient composition, will affect development, growth, reproductive capacity, ?immunocompetence,? and disease pathology of Zebrafish and their offspring in a standard and defined manner. This hypothesis is based on the fact that different commercial diets currently referenced in Zebrafish studies elicit different survival and growth rates within specific Danio rerio cohorts. Feeding different commercial diets also causes differences in reproductive success, indicating that one or more physiological functions are affected by the nutrient composition of the diet. Further, these commercial diets are not an open formulation and thus may change without the knowledge of the scientist thereby his/her research may become unverifiable and not repeatable. Also, the lack of control of diet influences other specific behavioral, physiological, biochemical, or molecular pathways, leading to a loss of rigor and reproducibility. As a consequence, undesired variability in the outcome of specific experimental trials designed to address developmental, pharmacological, toxicological, or disease-related questions have emerged, confounding progress in this field. Our mutual Milestones and Specific Experimental Aims are straightforward. Milestone 1 seeks the development and optimization of candidate reference diets for Zebrafish, based largely on previous work in the lab at UAB. Specific Aims for Milestone 1 will (a) test a small series of candidate diets that differ modestly in composition and (b) continue to develop specific outcome assessment tools to evaluate nutrient effects on organismal, physiological, and molecular levels of biological organization. A lead candidate diet that is chemically defined and easy to manufacture will emerge from these initial studies. Additional back-up lead diets will also advance into Milestone 2 efforts below. Early juvenile and adult Danio rerio will be evaluated. We will compare our diet throughout the study to other diets used currently in Zebrafish husbandry. Milestone 2 seeks to select of a Standard Reference Diet for Zebrafish. Specific Aims for Milestone 2 will (a) test coded and blinded lead and back-up lead diets in 3 independent Zebrafish facilities across the US for rigor, reproducibility and independent validation of the data and (b) develop assay metrics led by our laboratory among these Zebrafish facilities for quantitative and reproducible testing of diets. A standard reference diet is envisioned to emerge and be selected for further development in Phase II SBIR envisioned activities that will be spearheaded eventually by a new subsidiary of Meridian Biotech.